CN106492271B - The preparation of the antibacterial rush difunctional Guided Bone Regeneration Absorbable membrane of synosteosis - Google Patents
The preparation of the antibacterial rush difunctional Guided Bone Regeneration Absorbable membrane of synosteosis Download PDFInfo
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Abstract
The invention discloses a kind of preparation methods of Guided Bone Regeneration magnesium alloy film for promoting synosteosis function with antibacterial.This method embodiment includes magnesium alloy film, the operation followed the steps below to magnesium alloy film: prepares calcium phosphor coating in magnesium alloy film surface, realization slows down magnesium alloy membrane degradation speed.BMP-2 is loaded in the magnesium alloy film surface containing calcium phosphor coating；Nano-Ag particles are loaded in the magnesium alloy film surface for being mounted with BMP-2.This kind of new membrane can solve the shortcomings that existing Absorbable membrane can not be established preferably and maintain osteanagenesis space, the shortcomings that existing nonabsorable film needs second operation to take out, more importantly it can unsuccessfully provide a kind of effective solution approach to solve GBR postoperative infection and synosteosis.
Description
Technical field
The present invention relates to a kind of fields of biomedicine, more particularly, to a kind of guidance bone for promoting synosteosis function with antibacterial
The preparation method of recycling magnesium alloy film.
Background technique
Titanium or titanium alloy implantation has become the important restorative procedure of the various organ diseases of the mankind or defect missing, it is wide
The general fixed device of dental implant, maxillofacial prosthesis body spud pile, hearing aid/justice ear, joint prosthesis, the bone of being applied to is implanted into formula vacation
Many clinical fields such as limb, cardiac stent, joint prosthesis.However alveolar ridge atrophy caused by osteoporosis, severe periodontitis,
The diseases such as tumor post-operation, diabetes have limited to the utilization of this technology.In order to solve the demand of this some patients, bone increment is
Truly feasible solution.Artificial bone and Guided Bone Regeneration membrane material is clinically mostly used to carry out bone increment, however
Carrying out vertical and horizontal bone increment using Guided Bone Regeneration film and artificial bone, there are following difficult points: 1) modeling of repair materials
Shape 2) osteanagenesis space foundation and maintain 3) blood supply 4) prevention of postoperative infection 5) bone increment bone sclerotin.Magnesium alloy
With good Mechanical Property of Metal, in people's degradable in vivo, and have absorbability, it can effective ground heddle during being degraded due to it
It closes acidic materials and promotes pH, therefore there is certain antibacterial ability.This makes magnesium alloy be expected to become optimum Guided bone regeneration
Membrane material.But magnesium alloy materials have the following problems: 1) degradation speed is too fast, and can generate a large amount of hydrogen, can not be in long enough
Period in maintain osteanagenesis space 2) though magnesium alloy has certain antibacterial ability, however its anti-microbial property is still more weak,
When infecting more serious, guided bone regeneration (Guided bone regeneration, GBR) failure possibility is larger 3)
It promotes bone regeneration capability weaker.Therefore magnesium alloy film is modified make it have antibacterial and promote synosteosis ability can obtain it is good
Good effect.
Summary of the invention
The purpose of the present invention is overcome the deficiencies of the prior art and provide one kind in magnesium alloy film surface prepares coating to slow down
Its degradation speed, and load Bone Morphogenetic Protein-2 (Bone morphogenetic protein-2, BMP-2) in the coating and receive
Rice Argent grain makes it have antibacterial and promotes the preparation method of synosteosis function.
The invention adopts the following technical scheme: a kind of Guided Bone Regeneration magnesium alloy film for promoting synosteosis function with antibacterial
Preparation method, it is characterized in that including magnesium alloy film, the operation that magnesium alloy film is followed the steps below:
The first step polishes magnesium alloy film SiC abrasive paper for metallograph, cleaning, drying；
Second step prepares calcium phosphor coating in magnesium alloy film surface, and realization slows down magnesium alloy membrane degradation speed；
Third step loads BMP-2 in the magnesium alloy film surface containing calcium phosphor coating, has magnesium alloy film and promote synosteosis
Function；
4th step, be mounted with BMP-2 magnesium alloy film surface load nano-Ag particles, make magnesium alloy film have antibacterial and
Promote the function of synosteosis.
As an improvement the preparation of calcium phosphor coating is realized by biomimetic mineralization method, calcium phosphor coating in the middle second step
Use the simulation physiological fluid of 5 times of concentration with biomimetic mineralization method preparation, biomimetic mineralization method prepare calcium phosphor coating the following steps are included:
The preparation of the simulation physiological fluid of [1] 5 times of concentration: according to required liquid volume, keep ion concentration as follows: sodium
Ion 733.5mM, magnesium ion: 7.5mM, calcium ion 12.5mM, chloride ion 720.0mM, phosphoric acid hydrogen radical ion 5.0mM, sulfate radical
Ion 2.5mM, bicarbonate ion 21.0mM；
[2] pretreatment of magnesium alloy film: by magnesium alloy film with successively with p1000, the SiC abrasive paper for metallograph of p1800, p2000
Polishing is cleaned with acetone, dehydrated alcohol, deionized water, is dry under nitrogen stream；
[3] calcium phosphor coating is prepared in Mg alloy surface: magnesium alloy being soaked in the simulation physiological fluid of 5 times of concentration, is placed in
Temperature regulates and controls on heater, thermoreceptor probe is immersed in solution, 37 DEG C of constant temperature, bar magnet Stirring, 250 circles/min.
When the pH of solution is increased to 7.8 or more, reaction is terminated, takes out sample, excessive moisture is sucked out with paper, for use；
As an improvement the preparation mixed with the calcium phosphor coating of BMP-2 is prepared by BMP-2 and coating co-electrodeposition method, institute
It states in third step and is filled by co-electrodeposition method in magnesium alloy film surface with the over-saturation calcium phosphorus solution mixed with BMP-2 with calcium phosphor coating
Carry BMP-2 coating the step of the following steps are included:
[1] preparation of over-saturation calcium phosphorus solution: according to required liquid volume, keep ion concentration as follows: sodium ion
140.0mM, calcium ion 4.0mM, chloride ion 144.0mM, phosphoric acid hydrogen radical ion 2.0mM, and buffered with Tris liquid, supplement phase
The ion answered；
[2] it takes in a small amount of [1] prepared solution in EP pipe, and BMP-2 is added in the solution, mixing piping and druming, until
It is all dissolved, and then is fully transferred to fill in [1] in prepared reaction unit by the liquid, is mixed, is made its final concentration
For 0.1mg/ml；
[3] preparation of BMP-2 carrying layer: the magnesium alloy film for preparing calcium phosphor coating in second step is soaked in [2] and is matched
It in the over-saturation calcium phosphorus solution made, is placed in 37 DEG C of water bath with thermostatic control shaking tables, shaking table frequency is 60 times/min, after 48 hours eventually
It only reacts, discards over-saturation calcium phosphorus solution, prepare cated sample and be placed in lyophilized preparation, freeze-drying is stand-by overnight；
Realized as an improvement being loaded with the preparation of the calcium phosphor coating of nano silver by immersion-czochralski method, it is described in the 4th
Nano-Ag particles are loaded in step uses [Ag (NH3)2] to impregnate-czochralski method preparation, lifting-infusion method preparation is mounted with OH solution
The calcium phosphor coatings of nano-Ag particles the following steps are included:
[1] [Ag(NH3)2] OH solution preparation: the ammonium hydroxide of 2wt% is instilled dropwise in the silver nitrate solution of 2wt%, simultaneously
Ultrasonic vibration is carried out, until solution is become colorless clear solution by light brown；
[2] preparation of nano silver carrying layer: the sample prepared in third step is soaked in over-saturation calcium phosphorus solution, and
It is placed in 37 DEG C of water bath with thermostatic control shaking tables, shaking table frequency is 60 times/min, and reaction is terminated after 48 hours, it is molten to discard over-saturation calcium phosphorus
Liquid prepares cated sample and is placed in lyophilized preparation, and freeze-drying is overnight；The solution that the sample prepared is prepared in [1]
Middle immersion 20S, is then at the uniform velocity lifted with the speed of 1mm/s, is dried at 37 DEG C, after drying by sample be placed in closed container with
The glucose solution of 2wt% reacts 2 hours at 70 DEG C, dry under nitrogen stream；
Beneficial effects of the present invention: magnesium alloy film surface is loaded with the main spy of nano silver and the calcium phosphor coating coating of BMP-2
Sign has following three points: 1. the component and structure of coating change in continuity；2. material internal does not have sharp interface；3. the property of material
Matter also accordingly changes in gradient.Have the advantage that the coating because of features described above
(1) slow down its degradation speed by modifying calcium phosphor coating in Mg alloy surface, it can be in the sufficiently long time
Interior maintenance osteanagenesis space；
(2) nano silver and BMP-2 are loaded in calcium phosphor coating, make its GBR is postoperative can be with slow release nanometer silver to reach
Good antibacterial effect；Meanwhile the BMP-2 of loading can help preferably skeletonization, to promote the success rate of GBR；
(3) nano silver is a kind of good antibacterial agent, and some researches show that it can have good sterilization to imitate to various bacteria
Fruit, including fungi and some bacteriums for having drug resistance.In addition some researches show that nano silvers not to allow to be also easy to produce drug resistance, low concentration
Nano silver it is harmless and microbial reproduction, growth can be effectively inhibited or cause its death.Therefore, the antibacterial agent of silver is carried
It has been widely used in weaving, environmental protection, building materials and medical domain；
(4) Bone Morphogenetic Protein-2 (Bone morphogenetic protein-2, BMP-2) is that one kind is fixed by disulfide bond
Double-chain polypeptides dimer molecule, and passed through U.S. FDA authentication approval its carry out clinical application.A large amount of zoopery and face
Bed test confirms it with good Guided Bone Regeneration ability.The application of BMP-2 is mostly the side that BMP-2 is carried with polymer at present
Formula.Its major way is the surface that the growth factors such as BMP-2 are directly adsorbed in polymer.However it is this in the suction of polymer surface
Attached bone growth factor is only capable of forming limited, two-dimensional albumen memory space, when being exposed to physiological environment, growth factor meeting
It discharges rapidly, cannot achieve long-term slow control release, so that it is unable to maintain that the microenvironment of induced osteogenesis.In order to overcome this
A disadvantage, many products attempt to increase substantially bone growth factor dosage, and the dosage of some products is the mg grade of grade.Although
It solves the problems, such as to maintain induced osteogenesis microenvironment to a certain extent, but also brings many new problems and widely apply bon e formation
Albumen causes valuable product, influence its can be formed in a short time in clinical popularization and application high dose Bone Morphogenetic Protein it is high
Local concentration, will cause hyperfunction, bone adhesion of osteoclast function etc..Other loads that can be realized bone growth factor slow release
Body, such as microcapsules, microparticle etc. need to be related to during the preparation process the harmful substances such as organic solvent mostly, and at present
It is not possible in conjunction with polymer, it can by biomimetic mineralization method and the calcium phosphor coating for being mounted with BMP-2 with the preparation of coating co-electrodeposition method
To well solve problem above；
(5) this Absorbale magnesium alloy film with antibacterial and promotion osteogenic action that the present invention designs can be well
Solve some defects existing for existing inducting osseous tissue regeneration film；
(6) this kind of new membrane can solve existing Absorbable membrane and can not preferably establish and maintain lacking for osteanagenesis space
The shortcomings that point, existing nonabsorable film needs second operation to take out, more importantly it can for solve GBR postoperative infection with
And synosteosis unsuccessfully provides a kind of effective solution approach.
Detailed description of the invention: Fig. 1 is the structure chart for the magnesium alloy film that the present invention is mounted with nano silver and the calcium phosphor coating of BMP-2.
Wherein 1 is magnesium alloy film, and 2 be the calcium phosphor coating for being mounted with nano silver and BMP-2, from top to bottom respectively carrying nano silver layer, calcium
Phosphorous layer carries BMP-2 layers, calcium-phosphate layer.
Specific embodiment
It elaborates below in conjunction with attached drawing to specific embodiments of the present invention.
As shown in Figure 1, promoting the guidance bone of synosteosis function again with antibacterial for a kind of of preparation method that the present invention has
Raw magnesium alloy film specific embodiment.The embodiment includes magnesium alloy film, the operation followed the steps below to magnesium alloy film:
The first step polishes magnesium alloy film SiC abrasive paper for metallograph, cleaning, drying；
Second step prepares calcium phosphor coating in magnesium alloy film surface, prepares 5 times of concentration of analog physiological fluids, and take 10ml, leads to
Biomimetic mineralization method is crossed, prepares calcium phosphor coating in magnesium alloy film surface, realization slows down magnesium alloy membrane degradation speed；
Third step loads BMP-2 in the magnesium alloy film surface containing calcium phosphor coating, has magnesium alloy film and promote synosteosis
Function；
4th step, be mounted with BMP-2 magnesium alloy film surface load nano-Ag particles, make magnesium alloy film have antibacterial and
Promote the function of synosteosis；
Oneself warp of biomimetic mineralization, which becomes, in recent years increases one of most common method of polymer bone guided.But bionical mine
Change method also has disadvantage.The same with organism inner bone tissues mineralising, biomimetic mineralization must can just be opened by active group as nucleus
Dynamic calcium phosphorus precipitation.And polymer surface exactly lacks these active groups.Scholars are by the methods of hydrolysis and alkali process to more
Polymer surface is modified, and active group is added, to improve the ability of polymer spatial induction biomimetic mineralization.But these tables
The applicability of surface modification method is very limited, is unable to satisfy the needs in various polymer surfaces prepares coating, and polymer
Biocompatibility may also change in surface modification process.Therefore, biomimetic mineralization method is still by polymer surface
The limitations such as physics, chemical property induce the influence of mineralizer and can not be widely used.It finds one kind and does not depend on polymer surface
Property, the preparation method that can be widely applied for the bionic mineralization coat of a variety of polymers have great application value and meaning
Justice.The bionical calcium phosphor coating of two-part can in Mg alloy surface prepares coating and as the carrier of bioactie agent, realize because
The control release of son；
As an improvement specific embodiment, as an improvement, the preparation of calcium phosphor coating passes through biomimetic mineralization method
It realizes, calcium phosphor coating uses the simulation physiological fluid of 5 times of concentration with the preparation of biomimetic mineralization method, biomimetic mineralization in the middle second step
Method prepare calcium phosphor coating the following steps are included:
The preparation of the simulation physiological fluid of [1] 5 times of concentration: according to required liquid volume, keep ion concentration as follows: sodium
Ion 733.5mM, magnesium ion: 7.5mM, calcium ion 12.5mM, chloride ion 720.0mM, phosphoric acid hydrogen radical ion 5.0mM, sulfate radical
Ion 2.5mM, bicarbonate ion 21.0mM；
[2] pretreatment of magnesium alloy film: by magnesium alloy film with successively with p1000, the SiC abrasive paper for metallograph of p1800, p2000
Polishing is cleaned with acetone, dehydrated alcohol, deionized water, is dry under nitrogen stream；
[3] calcium phosphor coating is prepared in Mg alloy surface: magnesium alloy being soaked in the simulation physiological fluid of 5 times of concentration, is placed in
Temperature regulates and controls on heater, thermoreceptor probe is immersed in solution, 37 DEG C of constant temperature, bar magnet Stirring, 250 circles/min.
When the pH of solution is increased to 7.8 or more, reaction is terminated, takes out sample, excessive moisture is sucked out with paper, for use；
As an improvement the preparation mixed with the calcium phosphor coating of BMP-2 is prepared by BMP-2 and coating co-electrodeposition method, institute
It states in third step and is filled by co-electrodeposition method in magnesium alloy film surface with the over-saturation calcium phosphorus solution mixed with BMP-2 with calcium phosphor coating
Carry BMP-2 coating the step of the following steps are included:
[1] preparation of over-saturation calcium phosphorus solution: according to required liquid volume, keep ion concentration as follows: sodium ion
140.0mM, calcium ion 4.0mM, chloride ion 144.0mM, phosphoric acid hydrogen radical ion 2.0mM, and buffered with Tris liquid, supplement phase
The ion answered；
[2] it takes in a small amount of [1] prepared solution in EP pipe, and BMP-2 is added in the solution, mixing piping and druming, until
It is all dissolved, and then is fully transferred to fill in [1] in prepared reaction unit by the liquid, is mixed, is made its final concentration
For 0.1mg/ml；
[3] preparation of BMP-2 carrying layer: the magnesium alloy film for preparing calcium phosphor coating in second step is soaked in [2] and is matched
It in the over-saturation calcium phosphorus solution made, is placed in 37 DEG C of water bath with thermostatic control shaking tables, shaking table frequency is 60 times/min, after 48 hours eventually
It only reacts, discards over-saturation calcium phosphorus solution, prepare cated sample and be placed in lyophilized preparation, freeze-drying is stand-by overnight；
Realized as an improvement being loaded with the preparation of the calcium phosphor coating of nano silver by immersion-czochralski method, it is described in the 4th
Nano-Ag particles are loaded in step uses [Ag (NH3)2] to impregnate-czochralski method preparation, lifting-infusion method preparation is mounted with OH solution
The calcium phosphor coatings of nano-Ag particles the following steps are included:
[1] [Ag(NH3)2] OH solution preparation: the ammonium hydroxide of 2wt% is instilled dropwise in the silver nitrate solution of 2wt%, simultaneously
Ultrasonic vibration is carried out, until solution is become colorless clear solution by light brown；
[2] preparation of nano silver carrying layer: the sample prepared in third step is soaked in over-saturation calcium phosphorus solution, and
It is placed in 37 DEG C of water bath with thermostatic control shaking tables, shaking table frequency is 60 times/min, and reaction is terminated after 48 hours, it is molten to discard over-saturation calcium phosphorus
Liquid prepares cated sample and is placed in lyophilized preparation, and freeze-drying is overnight；The solution that the sample prepared is prepared in [1]
Middle immersion 20S, is then at the uniform velocity lifted with the speed of 1mm/s, is dried at 37 DEG C, after drying by sample be placed in closed container with
The glucose solution of 2wt% reacts 2 hours at 70 DEG C, dry under nitrogen stream.
Claims (3)
1. a kind of preparation method for the Guided Bone Regeneration magnesium alloy film for promoting synosteosis function with antibacterial, it is characterized in that including that magnesium closes
Golden film, the operation that magnesium alloy film is followed the steps below:
The first step polishes magnesium alloy film SiC abrasive paper for metallograph, cleaning, drying；
Second step prepares calcium phosphor coating in magnesium alloy film surface, and realization slows down magnesium alloy membrane degradation speed；
Third step loads BMP-2 in the magnesium alloy film surface containing calcium phosphor coating, magnesium alloy film is made to have the function for promoting synosteosis
Energy；
4th step loads nano-Ag particles in the magnesium alloy film surface for being mounted with BMP-2, magnesium alloy film is made to have antibacterial and promotion
The function of synosteosis；
The preparation of calcium phosphor coating realizes that the loading of BMP-2 with coating co-electrodeposition method by realizing, nano silver by biomimetic mineralization method
Loading realize that calcium phosphor coating uses the simulation physiological fluid of 5 times of concentration with bionical in the second step by immersion-czochralski method
The preparation of mineralising method, biomimetic mineralization method prepare calcium phosphor coating and comprise the steps of:
The preparation of the simulation physiological fluid of [1] 5 times of concentration: according to required liquid volume, keep ion concentration as follows: sodium ion
733.5mM, magnesium ion: 7.5mM, calcium ion 12.5mM, chloride ion 720.0mM, phosphoric acid hydrogen radical ion 5.0mM, sulfate ion
2.5mM, bicarbonate ion 21.0mM；
[2] pretreatment of magnesium alloy film: magnesium alloy film being used and successively uses p1000, the SiC abrasive paper for metallograph polishing of p1800, p2000,
It is cleaned with acetone, dehydrated alcohol, deionized water, is dry under nitrogen stream；
[3] calcium phosphor coating is prepared in Mg alloy surface: magnesium alloy being soaked in the simulation physiological fluid of 5 times of concentration, is placed in temperature
Regulate and control on heater, thermoreceptor probe immersed in solution, 37 DEG C of constant temperature, bar magnet Stirring, 250 circles/min,
When the pH of solution is increased to 7.8 or more, reaction is terminated, takes out sample, excessive moisture is sucked out with paper, for use.
2. a kind of preparation side of Guided Bone Regeneration magnesium alloy film for promoting synosteosis function with antibacterial according to claim 1
Method, it is characterized in that prepared mixed with the preparation of the calcium phosphor coating of BMP-2 by BMP-2 and coating co-electrodeposition method in the third step,
Pass through co-electrodeposition method in magnesium alloy film surface with calcium phosphor coating with the over-saturation calcium phosphorus solution mixed with BMP-2 in the middle third step
Load BMP-2 coating the step of the following steps are included:
[1] preparation of over-saturation calcium phosphorus solution: according to required liquid volume, keep ion concentration as follows: sodium ion 140.0mM,
Calcium ion 4.0mM, chloride ion 144.0mM, phosphoric acid hydrogen radical ion 2.0mM, and being buffered with Tris liquid, supplement accordingly from
Son；
[2] it takes in a small amount of [1] prepared solution in EP pipe, and BMP-2 is added in the solution, mixing piping and druming, until it is complete
Portion's dissolution, the liquid then be fully transferred to fill in [1] in prepared reaction unit, is mixed, is kept its final concentration of
0.1mg/ml；
[3] preparation of BMP-2 carrying layer: the magnesium alloy film for preparing calcium phosphor coating in second step is soaked in [2] and is prepared
Over-saturation calcium phosphorus solution in, be placed in 37 DEG C of water bath with thermostatic control shaking tables, shaking table frequency is 60 times/min, is terminated after 48 hours anti-
It answers, discards over-saturation calcium phosphorus solution, prepare cated sample and be placed in lyophilized preparation, freeze-drying is stand-by overnight.
3. a kind of preparation side of Guided Bone Regeneration magnesium alloy film for promoting synosteosis function with antibacterial according to claim 1
Method, it is characterized in that in load nano-Ag particles in the 4th step and use [Ag (NH3)2] OH solution with impregnate-czochralski method preparation, lifting-
Infusion method preparation be mounted with nano-Ag particles calcium phosphor coating the following steps are included:
[1][Ag(NH3)2] OH solution preparation: the ammonium hydroxide of 2wt% is instilled dropwise in the silver nitrate solution of 2wt%, at the same into
Row ultrasonic vibration, until solution is become colorless clear solution by light brown；
[2] preparation of nano silver carrying layer: the sample prepared in third step is soaked in over-saturation calcium phosphorus solution, is placed in
In 37 DEG C of water bath with thermostatic control shaking tables, shaking table frequency is 60 times/min, and reaction is terminated after 48 hours, discards over-saturation calcium phosphorus solution, is made
Standby cated sample is placed in lyophilized preparation, and freeze-drying is overnight；The sample prepared is impregnated in the solution that [1] prepares
Then 20S is at the uniform velocity lifted with the speed of 1mm/s, dry at 37 DEG C, is placed in sample in closed container and 2wt% after drying
Glucose solution reacted 2 hours at 70 DEG C, it is dry under nitrogen stream.
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| CN108295307A (en) * | 2018-03-02 | 2018-07-20 | 中国医科大学附属口腔医院 | A kind of bone implantation calcium phosphorous compound-magnesium composite coating and preparation method thereof |
| CN112675364A (en) * | 2021-01-05 | 2021-04-20 | 宁波瑞瑧生物科技有限公司 | Composite guided tissue regeneration membrane and preparation method thereof |
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